Transcript MADCAM1 GENE POLYMORPHISMS AND THE OUTCOME OF …
MADCAM1
GENE POLYMORPHISMS AND THE OUTCOME OF THE ALLOGENEIC HAEMATOPOIETIC STEM CELL TRANSPLANTATION
Z. Ambruzova 1 , F. Mrazek 1 , L. Raida 2 , A. Stahelova 1 , E. Faber 2 , K. Indrak 2 , M. Petrek 1 1 Laboratory of Immunogenomics and Proteomics, Department of Immunology, 2 Department of Haematooncology Palacky University and University Hospital Olomouc, Czech Republic
Supported by Czech Govt. Funding MSM 6198959205 and IGA MZ CR NR 9099
INTRODUCTION
various outcome
non-HLA gene polymorphisms
are intensively studied for their possible relevance for allogeneic haematopoietic stem cell transplantation (aHSCT)
migration and distribution of activated donor T cells
to the recipient mucosal sites and parenchymal target organs is important for development of graft-versus-host disease (GVHD) organ specific „homing“ of donor cells is mediated via interaction between
adhesion molecules
and their ligands
MUCOSAL ADDRESSIN CELL ADHESION MOLECULE-1
(
MAdCAM-1)
glycoprotein expressed on the surface of the endothelial cells (high level of expression in Peyer ´s patches and mesenteric lymph nodes)
ligands
: α4β7 integrins (LPAM-1, CD49d/β7) L-selectin (CD62L)
function
: lymphocyte traffic to mucosal sites Eksteen et al, Clin Med 2004; 4:173-80 adhesion of leucocytes on the endothelium aHSCT donor T cells „homing“ into the recipient mucosal tissue
MADCAM1
GENE POLYMORPHISM
many single nucleotide polymorphisms (SNPs) were described throughout the
MADCAM1
gene (chromosome 19) particular variants of the
MADCAM1
gene may affect structure and/or expression of MAdCAM-1 molecule SNPs selected to our study: rs 758502 T/C promotor rs 2302217 G/A exon 3 synonymous (Pro/Pro) rs 3745925 G/T exon 4 non-synonymous (His/Pro)
AIM OF THE STUDY
to investigate whether the selected are the risk factors for development of the serious complications after aHSCT
MADCAM1
gene SNPs
MADCAM1 gene SNPs
rs 758502 T/C rs 2302217 G/A rs 3745925 G/T acute or chronic GVHD overall survival
INVESTIGATED SUBJECTS
aHSCT pairs Age – median (range)
Patients Donors
Recipient gender
Female Male
Diagnosis
Acute leukaemia (AML, ALL) Chronic leukaemia (CML, CLL) Non-Hodgkin lymphoma Other
Conditioning regimen
Non-myeloablative Myeloablative
Donor HLA compatibility
Identical Mismatched
87
44 (18-61) 40 (18-69) 37 50 43 15 14 15 48 39 87 0
Donor type
Related Unrelated
Cell source
PBSC Bone Marrow
Acute GVHD
Grade 0-I Grade II Grade III Grade IV 70 17 86 1 53 23 4 8
Chronic GVHD
None 56 Limited 17 Extensive 14
METHODS
1.
Genotyping
Polymerase Chain Reaction with Sequence Specific Primers (PCR SSP) Primers designed according to the reference
MADCAM1
gene sequence: rs 758502 T/C rs 2302217 G/A rs 3745925 G/T
TT TC CC
MADCAM1
genotyping (rs758502)
2. Statistics
Conformity to the Hardy-Weinberg equilibrium: Chi-square test Differences between allele and genotype frequencies: Pearson logistic regression model Overall survival analysis: Kaplan-Meier, log-rank test and Cox regression analysis (SPSS software, version 15.0) ´s Chi squared test, Bonferroni correction for multiple comparisons, binary
MADCAM1 SNPs GENOTYPE FREQUENCIES IN PATIENTS AND DONORS
Patients
n = 87
Donors
n = 85
MADCAM1 T/C (rs758502)
Genotype CC Genotype TC Genotype TT
MADCAM1 G/A (rs2302217)
Genotype AA Genotype GA Genotype GG
MADCAM1 G/T (rs3745925)
0.43 (37) 0.47 (41) 0.10 (9) 0.26 (22) 0.51 (43) 0.24 (20) 0.53 (44) 0.37 (31) 0.10 (8) 0.35 (29) 0.40 (33) 0.24 (20) Genotype GG Genotype GT 0.64 (54) 0.33 (28) 0.77 (63)* 0.18 (15) Genotype TT 0.04 (3) 0.05 (4) ______________________________________________________________________ *
MADCAM1
(rs3745925) GG genotype frequency: p = 0.03, p corr >0.05 for comparison patients with donors
FREQUENCIES OF MADCAM1 IN PATIENTS WITH AND WITHOUT ACUTE GVHD GENOTYPES
MADCAM1 T/C (rs758502)
Genotype CC Genotype TC Genotype TT
Patients aGVHD+
n = 35 0.37 (13) 0.54 (19) 0.09 (3)
MADCAM1 G/A (rs2302217)
n = 34 Genotype AA Genotype GA Genotype GG 0.15 (5) 0.62 (21) 0.23 (8)
Patients aGVHD-
n = 49 0.45 (22) 0.43 (21) 0.12 (6)
p*
0.477
0.592
n = 48 0.31 (15) 0.46 (22) 0.086
0.23 (11) 0.948
MADCAM1 G/T (rs3745925)
n = 34 n = 48 Genotype GG Genotype GT 0.59 (20) 0.35 (12) 0.69 (33) 0.29 (14) 0.354
Genotype TT 0.06 (2) 0.02 (1) 0.367
___________________________________________________________________________ * comparison of particular homozygous genotype versus carriage of an alternative allele
FREQUENCY OF CHRONIC GVHD IN RECIPIENTS ACCORDING TO THEIR MADCAM1 rs2302217 GENOTYPES
p = 0.03
p corr = NS 70% 60% 50% 40% 30% 20% 10% 0% 64,7% 34,0% Genotype AA Genotype AG or GG
MULTIVARIATE ANALYSIS OF RISK FACTORS FOR OVERALL SURVIVAL AFTER aHSCT
Factor
aGVHD cGVHD
MADCAM1
rs2302217 AA genotype
p
0,00001 <0,00005 0,001
HR
4,35 5,7 2,99
Factors included to the analysis:
Diagnosis (malignancy vs. non-malignancy), type of the donor (related vs. unrelated), stem cell source, conditioning regimen, GVHD prophylaxis, donor-recipient gender combination (female donor in male recipient vs. others), aGVHD, cGVHD,
MADCAM1
gene variants
CONCLUSION
preliminary data suggest that aHSCT
MADCAM1
rs2302217 AA genotype in recipients may be associated with the risk of chronic GVHD and decreased overall survival after possible impact of
MADCAM1
SNP variants for aHSCT outcome has to be confirmed in substantially larger cohorts of donor-recipient aHSCT pairs
Dept. of Immunology, Medical Faculty Palacky University and University Hospital Olomouc
Prof. Martin Petrek Frantisek Mrazek Anna Stahelova Jana Onderkova Silva Zachova
Dept. of Haematooncology, Medical Faculty Palacky University and University Hospital Olomouc
Prof. Karel Indrak Ludek Raida Edgar Faber
Thank you for your attention…